Novel Multidimensional Liquid Chromatography Workflow with In-Loop Enzymatic Digests of Multiple Heart-Cuts for Fast and Flexible Characterization of Biotherapeutic Protein Variants.

Multidimensional liquid chromatography (mD-LC) is becoming a powerful tool for complete characterization of individual peaks and protein variants through separation methods such as nondenaturing ion exchange (IEC) or size-exclusion chromatography coupled to reversed-phase (RP) chromatography. The flexibility of commercially available and customized mD-LC systems is still limited in terms of enzymatic peak processing between chromatographic dimensions. In this regard, only a few column-immobilized proteases are available for detailed peak characterization by mD-LC coupled to mass spectrometry (mD-LC-MS). Here, we present a purpose-built and automated multiple heart-cutting mD-LC design with a novel analytical workflow involving in-loop enzymatic heart-cut digestion between the first-dimensional column and transfer to the second dimension before MS or MS/MS analyses. The setup facilitates the spike-in of any enzyme to multiple heart-cuts for multilevel analysis, for example, for peptide mapping, fragment generation, or deglycosylation, to reduce heterogeneity and provide maximum flexibility in terms of incubation time for optimal peak characterization. We demonstrate the application of IEC coupled to RP-LC-MS and automated in-loop deglycosylation and on-column reduction of an IgG antibody combined with upper hinge region cleavage for Fab generation. We further employ mD-LC-MS and mD-LC-MS/MS to assess post-translational modifications of a bispecific antibody and to support molecule selection by evaluating the best downstream purification strategy. The novel design and automated workflow of the mD-LC system described here offers enhanced flexibility for in-solution processing and real-time monitoring of multiple heart-cuts enabling streamlined characterization of unknown biotherapeutic charge and size variants.

A randomized cross-over trial investigating the neurocognitive effects of acute exercise on face recognition in children with autism spectrum disorder.

Difficulties in face recognition contribute to social-cognitive problems in autistic children. Evidence on behavioral interventions targeting this cognitive domain is limited. In non-autistic individuals, a single exercise session is known to elicit temporary benefits for several cognitive functions. Our study investigates whether acute aerobic exercise influences face recognition in autistic children. In a randomized order, 29 participants completed a 20-min moderately-intense cycling bout on an ergometer and a control condition. Before and after each condition, participants categorized Mooney faces and instruments during a computerized cognitive task. Simultaneously, the N170 component of event-related potentials and pupil size were recorded using electroencephalography and eyetracking, respectively. As indicated by a greater increase of reaction time in the exercise compared to the control condition, the results revealed impaired face recognition following aerobic exercise. This effect was accompanied by a lower decrease of the positive N170 amplitude and a trend towards a greater constriction of the pupil size in the exercise compared to the control condition. Our findings highlight the interplay of the physiological state and face recognition in autistic children. Exercise-induced impairments in this social-cognitive ability may be due to an interference with the learning effect that is typically seen for the structural encoding of faces.

Are Student Teachers' Overall Expected Emotions Regarding Their Future Life as a Teacher Biased Toward Their Expected Peak Emotions?

Having functional expected emotions regarding one's future life as a teacher is important for student teachers to maintain their motivation to choose a career as a teacher. However, humans show several biases when judging their emotional experiences. One famous bias is the so-called peak-end effect which describes the phenomenon that overall affective judgments do not reflect the average of the involved emotional experiences but the most intense and the most recent of the involved emotional experiences. Regarding student teachers' expected positive emotions, such a bias would be functional since their motivation to become a teacher is enhanced. However, regarding student teachers' expected negative emotions, such a bias would be dysfunctional since their motivation to become a teacher would be decreased. The aim of the present preregistered study was to examine whether student teachers' expected future teaching-related emotions show a peak-end effect. Student teachers viewed 14 common events that could part of a typical everyday routine of a teacher and rated their expected emotional pleasure and discomfort for each of the events. Afterward, they were asked to rate their overall expected emotional pleasure and discomfort when looking at their future professional life as a whole. Results showed that expected pleasure was much larger than expected discomfort regarding both overall, peak, and average ratings. No peak-end effect was observed for overall expected discomfort which reflected the average expected discomfort across events. By contrast, overall expected pleasure was biased toward expected peak pleasure experiences. These findings indicate that student teachers judge their expected overall affect in a functional way: realistically when dealing with negative emotions but through rose-colored glasses when dealing with positive emotions.

The promotion of functional expected teaching-related emotions through expressive writing.

The aim of the present preregistered study was to examine whether expressive writing can help teacher students to develop functional expected teaching-related emotions. In a variation of James W. Pennebaker´s expressive writing paradigm, 129 teacher students were randomly assigned to write on three consecutive days either about the future teaching-related events that personally trigger the greatest fear and joy (treatment group: n = 67) or about a walk in a forest and a city park (control group: n = 62). In both groups, expected teaching-related positive emotions increased and expected teaching-related negative emotions decreased with increased writing sessions. After the writing sessions, the treatment group reported a stronger change in their view about their future professional life as a teacher, a more active personal involvement with their future professional life, and an increased motivation to use expressive writing in the future. These results demonstrate that expressive writing is a promising tool to promote teacher students' expected teaching-related emotions.

Teacher Well-Being: Teachers' Goals and Emotions for Students Showing Undesirable Behaviors Count More Than That for Students Showing Desirable Behaviors.

Previous findings indicate that the goals of teachers and their experienced emotions when interacting with students play an important role for their well-being. However, studies on the psychological impact of events have shown that the impact of bad events is stronger than the impact of good events. Thus, it may be that teachers' goals and emotions for students showing undesirable behaviors (e.g., students who disrupt the class, do not finish their work, and have a negative attitude to learning) contribute more to their well-being than teachers' goals and emotions for students showing desirable behaviors (e.g., students who pay attention in class, do their work on time, and have a positive attitude to learning), a distinction that has not been made in previous research. To examine this question, we measured teachers' goals and emotions for students showing desirable and undesirable behaviors, and their affective, evaluative, occupational, and psychological well-being (N = 250). The results showed that teachers' well-being was relatively strongly related to their goals and emotions for students showing undesirable behaviors: The higher the goals and the more positive the emotions, the higher the reported well-being. By contrast, the goals and emotions for students showing desirable behaviors were unrelated to teachers' well-being. These results demonstrate that the principle of "bad is stronger than good" holds also for the influence of teachers' goals and emotions on their well-being.

Differential characteristics in the chemical composition of bananas from Tenerife (Canary Islands) and Ecuador.

The contents of moisture, protein, ash, ascorbic acid, glucose, fructose, total sugars, and total and insoluble fiber were determined in cultivars of bananas (Gran Enana and Pequeña Enana) harvested in Tenerife and in bananas (Gran Enana) from Ecuador. The chemical compositions in the bananas from Tenerife and from Ecuador were clearly different. The cultivar did not influence the chemical composition, except for insoluble fiber content. Variations of the chemical composition were observed in the bananas from Tenerife according to cultivation method (greenhouse and outdoors), farming style (conventional and organic), and region of production (north and south). A highly significant (r = 0.995) correlation between glucose and fructose was observed. Correlations of ash and protein contents tend to separate the banana samples according to origin. A higher content of protein, ash, and ascorbic acid was observed as the length of the banana decreased. Applying factor analysis, the bananas from Ecuador were well separated from the bananas produced in Tenerife. An almost total differentiation (91.7%) between bananas from Tenerife and bananas from Ecuador was obtained by selecting protein, ash, and ascorbic acid content and applying stepwise discriminant analysis. By selecting the bananas Pequeña Enana and using discriminant analysis, a clear separation of the samples according to the region of production and farming style was observed.

Statistical differentiation of bananas according to their mineral composition.

The concentrations of Na, K, Ca, Mg, Fe, Cu, Zn, and Mn were determined in banana cultivars Gran enana and Pequeña enana cultivated in Tenerife and in cv. Gran enana bananas from Ecuador. The mineral concentrations in the bananas from Tenerife and from Ecuador were clearly different. The cultivar did not influence the mineral concentrations except in the case of Fe. Variations according to cultivation method (greenhouse and outdoors) and farming style (conventional and organic) in the mineral concentrations in the bananas from Tenerife were observed. The mineral concentrations in the internal part of the banana were higher than those in the middle and external parts. Representation of double log correlations K-Mg and Zn-Mn tended to separate the banana samples according to origin. Applying factor and cluster analysis, the bananas from Ecuador were well separated from the bananas produced in Tenerife, and therefore, these are useful tools for differentiating the origin of bananas.